Figure 5.

Signalling connections of PAC1 receptor. mRNA (A,D) and protein (B,E) expression of bone morphogenic protein (BMP)2, BMP4, BMP6, BMP7, BMPR1, Smad1, Indian Hedgehog (IHH), Sonic Hedgehog (SHH), Patched (PTCH)1 and Gli1 in cortical bone. For RT-PCR and Western blot reactions, actin was used as controls. Signals optical density was measured and results were normalised to the optical density of controls. For panels (A,D) and (B,E) numbers below signals represent integrated densities of signals determined by ImageJ software. (C) Smad1 immunohistochemistry in bones. Original magnification was 60×. Scale bar: 5 µm. (F) Three point bending tests of long bones. (a) Value of the loading force at crack (F_m), and (b) the value of the bending deformation at crack (Δl_m) were demonstrated. Asterisks indicate significant (* p < 0.05) alteration of expression as compared to the respective control. Representative results of 3 independent experiments.